Clinical Analysis Of The Association Between ANP32E And Gastric Carcinoma Progression And The Function Analysis Of The Effects Of ANP32E On Cell Migration And Invasion

BackgroundGastric carcinoma(GC)is one of the most common malignancies and the second leading cause of cancer death in the world.Recurrence and metastasis of gastric cancer are still important factors that threaten the survival of patients and the economic burden of GC.Previous studies have shown that acidic nuclear phosphoprotein 32 kilodalton E(ANP32E)play an important role in regulating nervous system development,synapse formation,and H2 A.Z chromatin assembly.In tumors,ANP32 E was also found to be upregulated in a number of tumor tissues and was associated with tumor metastasis and poor prognosis.However,studies on the function of ANP32 E in gastric cancer have not been reported.Therefore,in this study,the expression level of ANP32 E in gastric cancer tissues and cell lines was analyzed and the relationship between ANP32 E and clinical outcomes of GC patients was studied.Forthermore,to investigate the biological function of ANP32 E could provide theoretical basis for research and development of molecular markers for the treatment and prognosis of gastric cancer patients.MethodsIn order to clarify the differences in the expression level of ANP32 E between gastric cancer tissues and adjacent normal tissues,gastric cancer cell lines and normal gastric mucosa epithelial cells,quantitative real-time PCR(qRT-PCR),Wester blot and immunohistochemistry were used in this study.In order to clarify the clinical significance of ANP32 E expression,Chi-square test was used to analyze the relationship between ANP32 E expression and clinicopathological features of gastric cancer.Kaplan-Meier regression was used to analyze the five-year survival time of gastric cancer for each pathological parameter.Cox proportional hazards model was used to analyze the risk factors of gastric cancer progression in these 4 factors.To clarify the role of ANP32 E in the metastasis of gastric cancer,a lentiviral vector was used to construct an ANP32 E overexpressing cell line and a shRNA-mediated ANP32 E suppressing cell line.Traswell assay was used to detect the effect of ANP32 E overexpression and inhibition on the migration and invasion of gastric cancer cells.The mRNA and protein expression of metastasis-associated E-cadherin and MMP-7 were detected by qRT-PCR and Wester blot.ResultsANP32E was significantly up-regulated in clinical samples of gastric cancer.Compared with normal human gastric epithelial cells(GES1),the expression of ANP32 E in 8 gastric cancer cells(NCI-N87,AGS,MKN45,MKN28,SGC7901,MGC803,HGC27,and KAT03)was also significant increased.Correlative analysis of clinical data showed that the expression level of ANP32 E was associated to the degree of gastric cancer infiltration and clinical stage.The frequency of ANP32 E positive expression in patients with stage III+IV gastric cancer was significantly higher than that of stage I+II patients.The frequency of ANP32 E positive expression in non-invaded serosa of gastric cancer significantly occurred in patients with serosal layer infiltration.The survival analysis showed that patients with low expression of ANP32 E had longer disease-free survival time than patients with high expression of ANP32 E.After overexpression and inhibition of ANP32 E in gastric cancer cells,would healing assay and transwell experiments showed that the migrated and invasive cells in the ANP32E-overexpressing gastric cancer group increased significantly,while inhibition of ANP32 E expression could significantly reduce the metastatic potential of gastric cancer cells.Meanwhile,the mRNA and protein levels of E-cadherin and MMP-7 were significantly decreased in ANP32E-overexpressing cells,while the mRNA and protein expressions of E-cadherin and MMP-7 were significantly up-regulated in ANP32E-inhibited cells.ConclusionsThe expression of ANP32 E was up-regulated in gastric cancer tissues and high expression of ANP32 E was related to the poor prognosis of gastric cancer patients.ANP32 E can promote the migration and invasion of gastric cancer cells,and its potential regulatory mechanism is related to the up-regulation of E-cadherin and MMP-7 expression.